KR20240001759A - Composition for improving skin conditions comprising lactic acid bacteria fermentate of rice germ extract - Google Patents

Abstract

The present invention relates to a composition for improving skin condition containing an extract of lactic acid bacteria-fermented rice germ, and more specifically, to a food composition containing an extract of lactic acid bacteria-fermented rice germ, which exhibits effects of improving skin wrinkles, improving skin elasticity, and improving skin moisturization, and pharmaceuticals. It relates to cosmetic compositions and cosmetic compositions.
The composition of the present invention containing lactic acid bacteria-fermented rice germ extract inhibits the increase in skin epidermal thickness caused by ultraviolet ray stimulation, inhibits the decrease in skin collagen, inhibits the expression of collagen decomposition enzymes, and increases the expression of filaggrin in keratinocytes, thereby causing skin wrinkles. It can be very useful in developing agents for skin improvement, skin elasticity, and skin moisturization.

Description

Composition for improving skin conditions comprising lactic acid bacteria fermentate of rice germ extract}

The present invention relates to a composition for improving skin condition containing an extract of lactic acid bacteria-fermented rice germ, and more specifically, to a food composition containing an extract of lactic acid bacteria-fermented rice germ, which exhibits effects of improving skin wrinkles, improving skin elasticity, and improving skin moisturization, and pharmaceuticals. It relates to cosmetic compositions and cosmetic compositions.

Recently, as the average lifespan has been extended due to the development of medical technology, the quality of life has improved, and the desire for a healthy and beautiful life has increased, interest in skin care and health is expanding. Accordingly, various beauty functional cosmetics have been developed for the purpose of maintaining healthy skin, and in particular, research on preventing, alleviating, and improving the formation of skin wrinkles is actively underway. In addition, there are recent limitations in the ability of cosmetic ingredients to reach the skin dermis and provide nutrients, and as there is a change in perception that ingestion of food can provide nutrients or functional ingredients to the skin to improve skin beauty, inner beauty foods Research to discover materials is also actively underway.

The skin protects the internal organs from stimulation from the external environment and plays an important role in maintaining biological homeostasis, such as temperature regulation. As this type of skin ages, it is accompanied by external deformations such as decreased skin elasticity, changes in skin color, and formation of skin wrinkles. This skin aging process is caused by a combination of intrinsic aging, which occurs naturally as we age, and extrinsic aging, photoaging caused by the surrounding environment, especially ultraviolet rays (UV). There are detailed differences in the mechanisms by which wrinkles are formed between the two types of aging, but in the skin, the proliferation and metabolism of epidermal cells are reduced, the bond between the epidermis and dermis is weak, so it is easy to be damaged even by small stimuli, and collagen synthesis is reduced and excessive wrinkles occur. Physiological changes such as increased expression of collagen-decomposing enzyme MMPs (Matrix Metalloproteinases) occur, causing the skin to become dry, and due to structural changes in the epidermal and dermal layers, the skin loses elasticity and appears saggy, and wrinkles gradually deepen. The way to delay and suppress skin aging is to delay natural aging and effectively block photoaging caused by ultraviolet rays, which are environmental factors.

Recently, the destruction of the ozone layer due to environmental pollution has increased the amount of ultraviolet rays, and as a result, research on photoaging is attracting attention (Dermatology and Therapy, 23(1): 31-47, 2010). Appearance characteristics such as roughening, loss of elasticity, and wrinkles are observed in photoaged skin, and the main research field of photoaging is about changes in skin wrinkles. Regarding the formation of skin wrinkles due to photoaging, many research results have been reported on changes in basic physiological metabolism such as synthesis, decomposition, and moisture content of collagen, a major component of the skin (Annals of the New York Academy of Sciences, 973: 31-43, 2002).

Collagen present in the dermal layer of the skin accounts for approximately 70-80% of the total dry weight of the skin and is known to control the elasticity of the skin along with elastin, an elastic fiber. In particular, ultraviolet rays increase the production of reactive oxygen species and the skin's enzymatic and non-enzymatic antioxidant defense system collapses, increasing collagen decomposition and reducing biosynthesis, resulting in a significant decrease in collagen in the dermal layer (Journal of Investigative Dermatology, 126(12): 2565-2575, 2006). Matrix metalloproteinases (MMPs) have an important effect on the collagen reduction, which is involved in the decomposition of the extracellular matrix and basement membrane. Research results have reported that the activity of the enzyme is increased by ultraviolet rays, and that by inhibiting it, the increase in skin thickness and wrinkle formation induced by ultraviolet rays are reduced (Journal of Investigative Dermatology, 120(1): 128-134, 2003) . Therefore, controlling MMP is an effective method for preventing and treating photoaging.

The stratum corneum exists in the outermost layer of the skin and is in direct contact with the external environment, so it plays an important barrier function in protecting our bodies from external physical and chemical stresses. This barrier function is maintained by epidermal homeostasis. Epidermal homeostasis maintains continuous skin barrier function by forming a skin barrier called the stratum corneum through terminal differentiation through growth division and cell migration of keratinocytes in the basal layer (Korean Journal) of Food Science and Technology, 43: 458-463, 2011).

As keratinocytes differentiate, they produce two factors that affect moisturization. First, during keratinocyte differentiation, the cell membrane is replaced by a structure called the cornified envelope. The keratinocyte membrane is a membrane structure in which several structural proteins, including loricrin (LOR), involucrin (INV), and filaggrin (FLG), are cross-linked by enzymes called transglutaminases (TGM). It provides skin protection against the external environment and at the same time protects the skin from keratin. Inhibits moisture evaporation within cells. Keratinocyte membrane structural proteins and TGM begin to be expressed upon differentiation of keratinocytes and are therefore used as differentiation markers (Nature Reviews Molecular Cell Biology, 6(4): 328-340, 2005). Therefore, keratin cell membrane and differentiation factors are used as indicators of moisturization. Additionally, during the differentiation process of keratinocytes, keratinocytes produce natural moisturizing factor (NMF) and retain the function of a skin barrier. The protein that is an important source for the production of natural moisturizing factors is FLG. FLG is decomposed into hydrophilic amino acids by caspase 14 to form natural moisturizing factors. Natural moisturizing factors function to maintain moisturizing power in the skin by providing water holding capacity and moisture absorption from the atmosphere (Journal of Cell Science, 122: 1285-1294, 2009). Therefore, maintaining an appropriate level of natural moisturizing factors in the skin is a very important factor for skin health through skin barrier function. Second, the interkeratinocyte lipids that make up the stratum corneum of the skin are composed of lipid components such as ceramide, cholesterol, and free fatty acids. Ceramide, a type of sphingolipid, accounts for more than 40% of lipids between keratinocytes and is an essential component that forms the structure of the stratum corneum that functions as a skin barrier. Ceramide synthase (CerS) is the most important enzyme in ceramide synthesis, and most of the ceramide present in the stratum corneum is synthesized by CerS3 (Biochimie, 91; 784-790, 2009; Biochimica et Biophysica Acta, 1841; 422 -434, 2014).

Accordingly, the present inventor has conducted repeated research to develop a substance derived from natural products that has the effect of preventing, improving, or treating skin wrinkles, loss of elasticity, and loss of moisture in the skin, and as a result, high content of γ-aminobutyric acid (GABA) was used. The present invention was completed after discovering that a lactic acid bacteria-fermented rice germ extract exhibits a very excellent effect in improving skin condition.

Therefore, the purpose of the present invention is to provide a food, pharmaceutical or cosmetic composition for improving skin elasticity or wrinkles containing lactic acid bacteria fermented rice embryo extract as an active ingredient.

Another object of the present invention is to provide a food, pharmaceutical or cosmetic composition for improving skin moisturization containing lactic acid bacteria fermented rice germ extract as an active ingredient.

In order to achieve the above-described object of the present invention, the present invention provides a food, pharmaceutical or cosmetic composition for improving skin elasticity or wrinkles containing lactic acid bacteria fermented rice embryo extract as an active ingredient.

In order to achieve another object of the present invention, the present invention provides a food, pharmaceutical or cosmetic composition for improving skin moisturization containing lactic acid bacteria fermented rice germ extract as an active ingredient.

Hereinafter, the present invention will be described in detail.

Throughout this specification, '%' used to indicate the concentration of a specific substance means (w/w) % for solid/solid, (w/v) % for solid/liquid, and Liquid/liquid is (v/v) %.

The term “unripe embryo extract” in the present invention includes an extract obtained by extracting and processing clean rice embryos produced in a processing plant equipped with an embryo selection system with a solvent such as water, and a dilution or concentrate of the extract.

The method for extracting the embryo is not particularly limited, and may be extracted according to a method commonly used in the art. Non-limiting examples of the extraction method include hot water extraction, ultrasonic extraction, filtration, and reflux extraction, which may be performed alone or by combining two or more methods.

The uncooked embryo extract of the present invention may be extracted after pre-treating the uncooked embryos, such as grinding, roasting, molding, extrusion, puffing, and superheated steam treatment, in order to stabilize the unborn embryos and increase extraction efficiency during extract production.

In one embodiment, the embryos are wetted to a moisture content of 10-20% by immersing or spraying water in water, and then treated with superheated steam at 170-350°C for 5-60 seconds in a fluidized bed device to achieve hydrothermal expansion. ) was processed.

The rice embryo extract of the present invention is prepared by mixing the pretreated rice embryo with one or more enzymes such as proteolytic enzymes, cellulolytic enzymes, and fat hydrolytic enzymes in order to increase extraction efficiency and produce various hydrolysis products during extract production. It may be an enzymatically hydrolyzed rice embryo extract extracted after hydrolysis with an enzyme.

In one embodiment, the embryo is ground to a 40-80 mesh and suspended in water at a concentration of 10%, and the proteolytic enzyme alcalase, the starch hydrolytic enzyme amylase, and the cellulolytic enzyme Cellucrast are added. (Celluclast) was added at an amount of 0.5 to 1.5% of the weight of each embryo, hydrolyzed at 50 to 70°C for 12 to 24 hours, and then extracted.

The rice embryo extract of the present invention is prepared by mixing the pretreated rice embryo with Bacillus subtilis TP-6 (KFCC 11343P), an excellent fermentation microorganism, in order to produce various bioactive substances such as functional peptides during extract production. ) It may be a fermented rice germ extract extracted after solid-state fermentation or deep fermentation.

The rice embryo extract of the present invention may be extracted using one or more substances selected from the group consisting of water and organic solvents having 1 to 8 carbon atoms. In the present invention, the extraction solvent may be a single or plural type of edible solvent, but is preferably water, alcohol, and alcohol having 1 to 6 carbon atoms [methanol (C1), ethanol (C2), propanol (C3), butanol ( C4), pentanol (C5), and hexanol (C6)] can be used. More preferably, in the present invention, the uncooked embryo extract may be obtained by extracting water with a solvent.

The rice embryo extract of the present invention can be used as a lactic acid bacteria medium after post-processing such as centrifugation, filtration, membrane separation, and chromatography.

In the present invention, before inoculating and fermenting lactic acid bacteria in the rice embryo extract, nutrients such as carbon source, nitrogen source, and minerals of the lactic acid bacteria and glutamic acid or glutamate are added to the rice embryo extract in order to promote the growth of the lactic acid bacteria and production of the desired metabolites. It can be added. The specific types of nutrients, their addition amounts, and fermentation conditions can be experimentally determined within the range of normal lactic acid bacteria fermentation known in the art, taking into account the intended fermentation time, degree of fermentation, etc.

The term “lactic acid bacteria fermented rice embryo extract” used in the present invention means that the rice embryo extract is fermented with lactic acid bacteria having GABA production ability. Optionally, after fermentation, products in the form of concentrates, powders, granules, crystals, etc. are obtained through several post-treatment processes such as sterilization, decolorization, filtration or centrifugation, concentration, ion exchange chromatography, crystallization, and drying. Formulated with is also included in the scope of the lactic acid bacteria fermented rice embryo extract.

In one embodiment, the lactic acid bacteria fermented rice germ extract may be a powder containing 10 to 20% (w/w) GABA. In one embodiment of the present invention, the fermented composition is a lactic acid bacteria-fermented rice embryo extract prepared by inoculating and fermenting the rice embryo extract with Lactobacillus sakei B2-16, accession number KFCC11321, as a fermentation source, and having a GABA of about 15. % (w/w).

In one embodiment, the lactic acid bacteria-fermented rice embryo extract may be purified and desalted to form a concentrate containing 20 to 80% (w/w) GABA.

In one embodiment, the fermented rice germ extract may be powder or granular containing 5 to 90% (w/w) GABA, preferably 10 to 20% (w/w).

In one embodiment, the lactic acid bacteria-fermented rice embryo extract may be in a crystal form with a purity of 90% or more after purification, desalting, concentration, and crystallization.

In the present invention, the rice germ (rice germ) is rich in nutrients such as proteins, vitamins, and minerals, and contains octacosanol, alpha-tocopherol, gamma oryzanol, gamma aminobutyric acid, dietary fiber, lecithin, and unsaturated fatty acids (linoleic acid, linolenic acid, It contains a large amount of excellent biologically active substances, including oleic acid, and is a safe and complete food and cosmetic material that cannot be compared to any other material.

Moreover, when various enzymatic hydrolysis, Bacillus subtilis, and lactic acid bacteria fermentation techniques are applied using the rice embryo as a fermentation source, various nutrients and bioactive substances derived from the rice embryo, as well as functional peptides and GABA produced during the fermentation process, are metabolized by useful microorganisms. Since the product will be produced, it will be possible to develop a fermentation product with a fusion concept that can simultaneously expect the effects of preventing, improving, or treating skin wrinkles, loss of elasticity, and loss of moisture in the skin.

According to one embodiment of the present invention, the lactic acid bacteria fermented rice germ extract has the effect of inhibiting the increase in skin epidermal thickness due to ultraviolet ray stimulation, inhibiting the decrease of skin collagen, inhibiting the expression of collagen decomposition enzyme, and increasing the expression of filaggrin in keratinocytes. It was confirmed to be very effective in improving skin wrinkles, skin elasticity, and skin moisturization.

In the present invention, “skin wrinkle improvement” means maintaining or strengthening the skin's ability to wrinkle and elasticity. Collagen, a collagen fiber in the dermal layer of the skin, and elastin, an elastic fiber, are the main proteins that play such roles and control skin elasticity, and the biosynthesis of collagen is affected by the internal and external influences of the skin. Specifically, in skin cells, when cell activity decreases due to natural aging, collagen fibers decrease, or active oxygen generated by excessive irradiation of ultraviolet rays or stress as an external factor destroys the thiol group (thiol: -SH) of the protein. It reacts with and inhibits enzyme activity or increases the expression of enzymes that decompose collagen, elastin, etc., increasing skin wrinkles and reducing elasticity, leading to skin aging.

In the present invention, “improvement of skin elasticity” refers to an increase in the elasticity of the skin, which inhibits collagen decomposition enzymes and promotes collagen production to suppress or inhibit the loss of skin elasticity or alleviate already reduced elasticity. says that

In the present invention, “promoting skin moisturization” refers to promoting the differentiation of keratinocytes to inhibit or suppress the decrease in skin moisture or to increase the moisture content of the skin to smooth the skin surface and provide gloss. says Additionally, it may mean that the expression of Filaggrin, a moisturizing protein contained in keratinocytes, is increased.

“Skin photoaging” in this specification refers to a physiological phenomenon in which the skin ages due to exposure to sunlight (ultraviolet rays). When exposed to the sun (ultraviolet rays) for a long period of time, deep wrinkles increase on the face and neck, and drying and drying of the skin occurs. It causes the skin surface to become rough or pigmentation such as spots and freckles.

The food composition of the present invention includes all types of functional foods, nutritional supplements, health foods, and food additives.

Food compositions of this type can be prepared in various forms according to conventional methods known in the art. Although not limited to this, for example, as a health food, the fermented lactic acid bacteria of fern extract can be prepared in the form of tea, juice, and drinks and consumed by liquefying, granulating, encapsulating, and powdering them for drinking. In addition, it can be prepared in the form of a composition by mixing the lactic acid bacteria fermented rice germ extract with known active ingredients known to be effective in improving skin condition. In addition, functional foods include, but are not limited to, beverages (including alcoholic beverages), fruits and their processed foods (e.g. canned fruit, bottled foods, jam, marmalades, etc.), fish, meat and their processed foods (e.g. ham, sausages, etc.) corned beef, etc.), bread and noodles (e.g. udon, buckwheat noodles, ramen, spaghetti, macaroni, etc.), fruit juice, various drinks, cookies, taffy, dairy products (e.g. butter, cheese, etc.), edible vegetable oil, margarine, vegetable oil It can be manufactured by adding lactic acid fermented rice germ extract to proteins, retort foods, frozen foods, and various seasonings (e.g., soybean paste, soy sauce, sauce, etc.). Additionally, in order to use the lactic acid bacteria fermented rice germ extract in the form of an additive, it can be prepared and used in the form of powder or concentrate.

The preferred content of the lactic acid bacteria fermented rice germ extract in the food composition of the present invention is not limited thereto, but is preferably 0.1 to 90% by weight of the final manufactured food. More preferably, the food composition containing the lactic acid bacteria fermented rice germ extract of the present invention as an active ingredient can be prepared in the form of a health functional food or dietary supplement by mixing it with an active ingredient known to have an effect of improving skin condition. there is.

The cosmetic composition of the present invention may be in the form of a general emulsified formulation or solubilized formulation. For example, lotion such as flexible lotion or nourishing lotion, emulsion such as facial lotion, body lotion, cream, essence, cosmetic ointment, spray, gel, pack, sunscreen, makeup base, liquid such as nutritional cream, moisture cream, eye cream, etc. It may have a formulation such as foundation type, solid type or spray type, powder, makeup remover such as cleansing cream, cleansing lotion, cleansing oil, cleansing foam, soap, body wash, etc. The cosmetic composition may further include commonly used ingredients depending on the formulation.

In addition, the cosmetic composition contains, in addition to the lactic acid bacteria fermented rice germ extract, fatty substances, organic solvents, solubilizers, thickening agents and gelling agents, softeners, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, Commonly used in water, ionic or non-ionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or in cosmetics. It may contain auxiliaries commonly used in the field of cosmetology, such as any other ingredients.

The cosmetic composition may contain a relatively high concentration of the Marian plum extract in the case of wash-off type cosmetics such as makeup removers and detergents in which the active ingredient stays on the skin for a short period of time. On the other hand, leave-on type cosmetics such as lotions, emulsions, creams, and essences, where the active ingredients stay on the skin for a long period of time, contain the Marian plum extract at a lower concentration than wash-off type cosmetics. It would be okay to do so. Although not limited thereto, in one embodiment of the present invention, the composition may include the Marian plum extract in an amount of 0.001% by weight to 10% by weight (preferably 0.01% by weight to 5% by weight) based on the total weight of the composition. there is. If the composition of the present invention contains the Marian plum extract in an amount of less than 0.001% by weight, a sufficient effect of preventing or improving skin aging, skin wrinkles, loss of elasticity and moisture loss in the skin cannot be expected, and if the composition of the present invention contains the Marian plum extract in an amount exceeding 10% by weight. This is to prevent unwanted reactions such as allergies or skin safety issues.

In addition, the cosmetic composition of the present invention can be provided as a cosmetic product having any one formulation selected from the group consisting of lotions, essences, skins, lotions, creams, and packs containing lactic acid bacteria fermented rice embryo extract as an active ingredient. .

Examples of the cosmetics include skin, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutritional lotion, massage cream, nutritional cream moisture cream, hand cream, foundation, essence, nutritional essence, pack, arsenic, cleansing. These include, but are not limited to, foam, cleansing lotion, cleansing cream, body lotion, and body cleanser.

The pharmaceutical composition according to the present invention can be formulated in various ways depending on the route of administration by methods known in the art along with a pharmaceutically acceptable carrier. The carrier includes all types of solvents, dispersion media, oil-in-water or water-in-oil emulsions, aqueous compositions, liposomes, microbeads and microsomes.

The pharmaceutical composition according to the present invention can be administered to a patient in a pharmaceutically effective amount, that is, an amount sufficient to exhibit the effects of improving skin wrinkles, improving skin elasticity, and improving skin moisturization. For example, a general daily dose may be administered in the range of 1 to 10,000 mg, preferably in the range of 10 to 200 mg. The pharmaceutical composition of the present invention can be administered once or in divided doses within a preferred dosage range. Additionally, the dosage of the pharmaceutical composition according to the present invention can be appropriately selected by a person skilled in the art depending on the administration route, administration subject, age, gender, weight, individual differences, and disease state.

The route of administration may be oral or parenteral. Parenteral administration methods include, but are not limited to, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual, rectal, or pancreatic. It may be administered internally, but is not limited to this, and most preferably, it may be administered orally.

When the pharmaceutical composition of the present invention is administered orally, powder, granules, tablets, pills, sugar-coated tablets, capsules, solutions, gels, syrups, suspensions, and wafers are prepared according to methods known in the art along with a suitable carrier for oral administration. It can be formulated in a form such as: Examples of suitable carriers include sugars including lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol and maltitol, starches including corn starch, wheat starch, rice starch and potato starch, cellulose, Fillers such as cellulose, including methyl cellulose, sodium carboxymethylcellulose, and hydroxypropylmethylcellulose, gelatin, polyvinylpyrrolidone, etc. may be included. Additionally, in some cases, cross-linked polyvinylpyrrolidone, agar, alginic acid, or sodium alginate may be added as a disintegrant. Furthermore, the pharmaceutical composition may further include anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers, and preservatives.

In addition, when administered parenterally, the pharmaceutical composition of the present invention can be formulated with a suitable parenteral carrier in the form of injections, transdermal administration, nasal inhalation, etc. according to methods known in the art.

Additionally, the pharmaceutical composition can be administered by any device that allows the active agent to move to target cells. Preferred administration methods and formulations include intravenous injection, subcutaneous injection, intradermal injection, intramuscular injection, or drip injection. Injections include aqueous solvents such as physiological saline solution or Ringer's solution, non-aqueous solvents such as vegetable oil, higher fatty acid esters (e.g., ethyl oleate, etc.), and alcohols (e.g., ethanol, benzyl alcohol, propylene glycol, or glycerin, etc.). It can be manufactured using stabilizers to prevent deterioration (e.g., ascorbic acid, sodium bisulfite, sodium pyrosulfite, BHA, tocopherol, EDTA, etc.), emulsifiers, buffers for pH adjustment, and agents to prevent microbial growth. It may contain pharmaceutical carriers such as preservatives (e.g., phenylmercuric nitrate, thimerosal, benzalkonium chloride, phenol, cresol, benzyl alcohol, etc.).

The above injections must be sterilized and protected from contamination by microorganisms such as bacteria and fungi. For injections, examples of suitable carriers include, but are not limited to, solvents or dispersion media including water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, etc.), mixtures thereof, and/or vegetable oils. You can. More preferably, suitable carriers include Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) containing triethanol amine, or isotonic solutions such as sterile water for injection, 10% ethanol, 40% propylene glycol, and 5% dextrose. etc. can be used. In order to protect the injection from microbial contamination, it may additionally contain various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc. Additionally, in most cases, the injection may additionally contain an isotonic agent such as sugar or sodium chloride.

As for other pharmaceutically acceptable carriers, those described in the following literature may be referred to (Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, PA, 1995).

The pharmaceutical composition according to the invention may contain one or more buffers (e.g. saline or PBS), carbohydrates (e.g. glucose, mannose, sucrose or dextran), antioxidants, bacteriostatic agents, chelating agents (e.g. For example, EDTA or glutathione), adjuvants (e.g. aluminum hydroxide), suspending agents, thickening agents and/or preservatives may be further included.

Additionally, the pharmaceutical compositions of the present invention can be formulated using methods known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal.

Additionally, the pharmaceutical composition of the present invention can be administered alone or in combination with known substances effective in improving skin condition.

The composition of the present invention containing lactic acid bacteria-fermented rice germ extract inhibits the increase in skin epidermal thickness caused by ultraviolet ray stimulation, inhibits the decrease in skin collagen, inhibits the expression of collagen decomposition enzymes, and increases the expression of filaggrin in keratinocytes, thereby causing skin wrinkles. It can be very useful in developing agents for skin improvement, skin elasticity, and skin moisturization.

Figure 1 is a graph showing the change in body weight of each animal group during the test period.
Figure 2 shows the results of observing the skin wrinkles of animals in the normal group (Normal), the ultraviolet irradiation group (UVB), and the test substance administration group after ultraviolet irradiation (FRGE) using replicas at 4, 8, and 12 weeks.
Figure 3 is a graph showing the results of quantifying the Total wrinkle area, Number of wrinkles, Total length, Mean length, and Mean depth of each animal group through replica analysis.
Figure 4 shows the results of quantification of redness, hydration, and thickness (caliper) in the skin of each animal group observed at 4, 8, and 12 weeks.
Figure 5 shows the results of quantification of skin elasticity observed in the skin of each animal group at 4, 8, and 12 weeks.
Figure 6 shows the results of H&E staining of skin tissue of each animal group and observation under a microscope after completion of the experiment.
Figure 7 shows the results of Masson trichrome staining of skin tissue of each animal group and observation under a microscope after completion of the experiment.
Figure 8 is a graph quantifying the expression level of MMP-3 mRNA observed by PCR in the skin tissue of each animal group after the end of the experiment.
Figure 9 is a graph observing and quantifying the expression level of filaggrin protein in skin tissue of each animal group by Western blotting after the end of the experiment.

Hereinafter, the present invention will be explained in detail by the following examples. However, the following examples are only for illustrating the present invention, and the present invention is not limited thereto.

Experiment method

1. Preparation of rice embryo extract

The moisture of the unborn embryos purchased from GoldenEye Co., Ltd. was adjusted to 20%, then treated with superheated water vapor at 290°C for 20 seconds in a vortex-type fluidized bed device, and then instantly released and cooled with room temperature air. 25 g of unexpanded embryos treated with superheated water vapor as described above and 10 times the amount of water were added to a 500 mL Erlenmeyer flask, and the top was sealed with aluminum foil.

Other unembryo samples were pulverized using a laboratory disk-type grinder, and then 25 g of the fraction passed through a 40-mesh sieve was suspended in 10 times the amount of water, and then alcalase, amylase, and celluclast were removed. Each was added singly or in combination at 1% of the ground embryo.

The superheated steam-treated US embryo suspension and the US embryo suspension flask to which hydrolytic enzymes were added after pulverization were extracted for 18 hours while stirring in a 55°C water bath, centrifuged at 13000 rpm for 15 minutes to obtain a supernatant, and the supernatant was filtered through a 0.2㎛ syringe filter. After filtration, the solid content in the supernatant was measured using a drying method at 105°C.

As shown in Table 1, the extraction yield was improved by more than 2 times by instantaneously hydrothermally expanding the embryo through superheated steam treatment, and the extraction yield was also increased by about 1.5 to 2 times with various enzymatic hydrolysis treatments, showing that it is effective as a pretreatment. You can.

[Table 1]

2. Preparation of lactic acid bacteria fermented rice embryo extract sample

Using a fermentation production facility, powdered samples of lactic acid bacteria fermented rice embryo extract were produced for animal testing. First, prepare 350L of 10 embryo extract medium in a 500L fermenter and add Lactobacillus pre-cultured in the laboratory. sakei B2-16 strain (accession number KFCC11321) culture medium was inoculated and cultured at 30°C for 48 hours. After completion of fermentation, the fermented broth was sterilized and filtered, dextrin was added, spray dried in a large spray dryer, and small packages of 1 kg were placed in aluminum laminated packaging containers.

As a result of analyzing the general components of the powder sample, calories (Kcal/100g) 261.3, ash (%) 28.5, carbohydrates (%) 45.9, moisture (%) 6.6, crude fat (%) 0.3, sodium (mg/100g) 3574.3, and The crude protein (%) was found to be 18.7, and the GABA content was confirmed to be 15% (w/w) through in-house analysis and by requesting analysis from the Korea Functional Food Research Institute.

3. Animal testing

(1) Experimental animals, breeding of experimental animals, test substances, and wrinkle-inducing

The mouse used in this experiment is a 6-week-old hairless mouse that is widely used in skin-related tests and is specific pathogen-free (SPF) of the HR-1 strain, which easily causes wrinkles and is congenitally hairless. ) were purchased from Joongang Laboratory Animal Co., Ltd. (Seoul, Korea) and allowed to acclimate for 7 days. During the acclimatization period, general symptoms were observed daily and only healthy animals were used in the experiment. The experimental animals were divided into three groups and each group was assigned 10 animals. The weight of each experimental animal was measured the day before the start of administration, and the ranked weights were used to separate groups into groups using a random method. For individual identification, the tail marking method and individual identification card marking method for each breeding box were used.

Feed and water were allowed to be consumed freely during the breeding period, and the temperature of the animal room was maintained at 23±3°C, relative humidity was 55±15%, and ammonia concentration was below 5ppm, and the light/dark cycle was 12 hours with fluorescent lighting (lights on at 08:00 ~ 20:00 (lights off). The illumination level of the breeding room was maintained at 268 Lux, the noise level at 55.1 dB, and the ammonia concentration at 5 ppm or less. Experimental animals were housed in stainless steel mesh breeding boxes (200W*260D*130Hmm) during the acclimatization, quarantine, administration, and observation periods.

The experimental group was divided into a normal group (Normal), an ultraviolet irradiation group (UVB), and a group that consumed lactic acid bacteria fermented rice germ extract (FRGE) along with ultraviolet irradiation. The test substances were mixed into the feed for each group and administered orally as solid content. The lactic acid bacteria fermented rice germ extract consumption group (FRGE) was administered a solid diet to which 100 mg (body weight/day/mouse) of the lactic acid bacteria fermented rice embryo extract powder sample prepared by the method of Experiment 2 above was added.

During the experimental breeding period, the backs of hairless mice were irradiated with ultraviolet rays (UV) three times a week using Philips TL20W/12. The amount of ultraviolet irradiation was measured at each irradiation using a Walkmann UV meter. The irradiation dose started at 1 MED (80 mJ/cm2) in the first week and increased by 1 MED every week. After 4 weeks, 4 MED was administered, and the irradiation dose was adjusted to ensure skin condition and appropriate wrinkle formation. A total of 80 MED of ultraviolet rays were irradiated.

(2) Test items

① Observation of general symptoms

All animals were observed for general symptoms once a day. On the day of UV irradiation, observations were made immediately after irradiation and every hour for 6 hours thereafter. General symptom observation continued until the end of the test.

②Weight measurement

Body weight was measured once a week (Monday) at a certain time (03:00 PM) for all animals.

③ Stereoscopic microscope photography

Stereomicroscopic images of the back skin area were taken using SMZ1500 (Nikon Japan) at 4, 8, and 12 weeks.

④ Production of copy version

A disk with a hole was attached to the back of a hairless mouse, a silicone rubber (Silflo: Flexico Developments Ltd, England) reagent was mixed, spread thinly on the surface of the disk, dried completely, and then carefully removed to create a replica. . Simulation plates were produced at weeks 4, 8, and 12, and the area, number, total length, average length, and average depth of wrinkles were analyzed using a skin wrinkle measuring device.

⑤ Measurement of moisture content, elasticity, and erythema

Skin moisture content and elasticity were measured in the same area as the replica production area. Skin moisture content was measured at 4, 8, and 12 weeks using a Corneometer (Courage and Khazaka, Germany).

Skin elasticity was measured at 4, 8, and 12 weeks using Cutometer (Courage and Khazaka, Germany). R0~9 and F0~1 were analyzed by repeating 1 second measurement at a sound pressure of 50 mbar and 1 second after inhalation three times.

Erythema caused by ultraviolet irradiation was measured with a spectrophotometer (KONICA MINOLTA, Tokyo, Japan).

⑥ Biopsy

At the end of the test, the skin on the back was removed and fixed in formalin. After step-by-step dehydration with alcohol and xylene, embedding in paraffin, sections of less than 5㎛ were made using a microtome, and the paraffin was removed again with alcohol and xylene.

Hematoxylin & Eosin (H&E) staining was performed and epidermal thickness was measured using NIS-Elements (Nikon, Japan). Additionally, Masson trichrome staining was performed to measure collagen production.

⑦ MMP-3 and filaggrin analysis

Skin tissue was extracted, RNA was extracted, synthesized into cDNA, and the expression level of MMP-3 was analyzed using RT-PCR. In addition, proteins were extracted from skin tissue and filaggrin expression levels were analyzed using western blotting.

Experiment result

1. General symptoms and changes in weight (Figure 1)

No unusual clinical symptoms due to oral administration of test substances were observed in any test system during the test period. Body weight also showed a normal increase, with no statistically significant difference (Figure 1).

2. Changes in skin wrinkle formation in hairless mice (Figures 2 and 3))

To evaluate the effect of ingestion of lactic acid bacteria-fermented rice germ extract on the degree of skin wrinkle formation, the area, number, and total number of wrinkles were measured at 4, 8, and 12 weeks using the naked eye and replicas of the back skin of experimental animals. Length, average length, and average depth were measured and statistically processed.

When observed with the naked eye during the 12-week UV irradiation period, the UVB control group repeatedly exposed to ultraviolet rays was observed to have numerous thick and deep wrinkles and an abnormal increase in the formation of fine wrinkles compared to the non-irradiated normal group (Figure 2) ). It was confirmed that the thickness and depth of wrinkles in the lactic acid bacteria fermented rice germ extract consumption group (FRGE) were significantly improved compared to the UVB group (Figure 2).

As a result of objectively evaluating the total area, total number, total length, average length, and average depth of wrinkles through simulation plate analysis, the FRGE intake group showed overall significant signs related to wrinkle formation and wrinkles compared to the UVB group (p< 0.05) (Figure 3).

3. Erythema, skin hydration and skin thickness (Figure 4)

In the 4th, 8th, and 12th weeks of the experiment period, the FRGE intake group showed a significant decrease in erythema and skin thickness compared to the UVB group (p<0.05).

And the skin hydration content in the FRGE intake group significantly increased compared to the UVB group (p<0.05).

4. Measurement of skin elasticity (Figure 5)

Skin elasticity was measured using a Cutometer at 4, 8, and 12 weeks after oral administration of the test substance. Skin is a viscoelastic material, and its deformation occurs through a combination of elastic deformation and viscous deformation. R2 (=Ua/Uf, total viscoelasticity) is a value that indicates how much the skin (Uf) that is maximally pulled up by the probe can return to its original state (Ua) at 10 seconds of suction. The closer it is to 1 (100%), the more viscoelastic it is. This means that R5 (=Ur/Ue, net elasticity) is the ratio of the skin height (Ue) lifted immediately after being suctioned by the probe and the skin height (recovered elasticity) returned immediately after the skin was lifted by opening the suction. Part) As a ratio of (Ur), the closer it is to 1 (100%), the more elastic the skin is. R7 (=Ur/Uf, recovery rate) is the proportion of the elastic part at the maximum skin height in 10 seconds. It is a value that indicates how much it occupies.

As a result of measuring skin elasticity, after 8 and 12 weeks in the FRGE intake group, the R2, R5, and R7 values were all significantly higher than those in the UVB group (p<0.05), showing that the skin elasticity decreased by ultraviolet irradiation was recovered. .

5. Histological evaluation (Figure 6, Figure 7)

To confirm histopathological changes on the last day of oral administration of the test substance, tissue analysis was performed after staining with hematoxylin and eosin (H&E). As a result of histopathology, it was observed that the thickness of the epidermal layer of the skin in the UVB group was significantly thickened compared to the normal group that did not receive ultraviolet irradiation. When photoaging progresses due to ultraviolet irradiation, etc., the formation of the stratum corneum increases to protect the dermal layer of the skin, making the skin thicker. Therefore, a thicker epidermal layer means that the skin damage caused by photoaging is greater. Meanwhile, the FRGE group that consumed the lactic acid bacteria-fermented rice germ extract had a significant decrease in epidermal thickness, which was almost similar to the Normal group, indicating that the tissue changes caused by photoaging were restored by the intake of the lactic acid bacteria-fermented rice embryo extract (Figure 6).

As a result of Masson Trichrome staining to confirm collagen in the dermal layer, it was confirmed that the UVB group had less collagen as staining loss was observed in the upper dermal layer compared to the Normal group, and no dermal layer damage was observed in the FRGE intake group. It was confirmed that collagen was restored by consuming lactic acid bacteria fermented rice germ extract (Figure 7).

6. Comparison of MMP-3 mRNA expression level using RT-PCR method (Figure 8)

The results of measuring the mRNA expression level of the MMP-3 enzyme gene, which is activated by UVB irradiation and decomposes collagen such as type IV, in skin tissue using RT-PCR are shown in Figure 8. The UVB group showed a significant increase in MMP-3 expression compared to the Normal group. The FRGE intake group showed significantly lower MMP-3 expression levels than the UVB group (p<0.05), indicating that intake of lactic acid bacteria fermented rice embryo extract had an effect of inhibiting the decomposition of extracellular matrix proteins.

7. Measurement of filaggrin using Western blot method (Figure 9)

The expression level of filaggrin, a moisturizing protein contained in keratinocytes forming the skin barrier, was measured using Western blot. In the UVB group, filaggrin expression decreased compared to the Normal group, and in the FRGE intake group, filaggrin expression significantly increased compared to the UVB group (p<0.05).

As a result of the above experiment, it was significantly confirmed that repeated oral administration of lactic acid bacteria fermented rice germ extract had the effect of suppressing wrinkle formation, moisturizing, and restoring elasticity to the skin caused by ultraviolet rays.

The composition of the present invention containing lactic acid bacteria-fermented rice germ extract inhibits the increase in skin epidermal thickness caused by ultraviolet ray stimulation, inhibits the decrease in skin collagen, inhibits the expression of collagen decomposition enzymes, and increases the expression of filaggrin in keratinocytes, thereby causing skin wrinkles. It can be very useful in developing agents for skin improvement, skin elasticity, and skin moisturization, so its industrial applicability is very high.

Claims (14)

A food composition for improving skin elasticity or wrinkles containing lactic acid bacteria fermented rice germ extract as an active ingredient.
A food composition for improving skin moisturization containing lactic acid bacteria fermented rice germ extract as an active ingredient.
The method of claim 1 or 2, wherein the lactic acid bacteria fermented rice embryo extract is powder or granular containing 5 to 90% (w/w) of GABA (γ-aminobutyric acid, GABA), or 20 to 80% (w) of GABA. /w) A food composition characterized in that it is one selected from the group consisting of a concentrate containing GABA and crystals containing 90% to less than 100% (w/w) GABA.
The food composition according to claim 1 or 2, wherein the lactic acid bacteria fermented rice embryo extract is obtained by fermenting the rice embryo extract with Lactobacillus sakei B2-16 (accession number KFCC11321).
The food composition according to claim 4, wherein the ungerm extract is extracted with one or more solvents selected from the group consisting of water, alcohols having 1 to 8 carbon atoms, and mixed solvents thereof.
The food composition according to claim 1 or 2, wherein the rice embryo extract is an extract of a rice embryo hydrothermally expanded with superheated steam, a rice embryo hydrolyzed by enzymes, or a combination thereof.
The food composition according to claim 6, wherein the enzyme is at least one selected from the group consisting of alcalase, amylase, and celluclast.
The method of claim 1 or 2, wherein the rice embryo extract is a rice embryo hydrothermally expanded with superheated steam, an enzymatically hydrolyzed rice embryo, or a combination thereof, Lactobacillus sakei B2-16 (accession number KFCC11321). ) A food composition characterized in that it is extracted after fermentation.
The method of claim 1 or 2, wherein the lactic acid bacteria fermented rice germ extract inhibits the increase in skin epidermal thickness caused by ultraviolet ray stimulation, inhibits the decrease in skin collagen, inhibits the expression of collagen degrading enzymes, and increases the expression of filaggrin in keratinocytes. A food composition characterized in that it exhibits at least one effect selected from the group consisting of.
The food composition according to claim 1 or 2, wherein the food composition is a health functional food.
A pharmaceutical composition for improving skin elasticity or wrinkles containing lactic acid bacteria fermented rice germ extract as an active ingredient.
A pharmaceutical composition for improving skin moisturization containing lactic acid bacteria fermented rice germ extract as an active ingredient.
A cosmetic composition for improving skin elasticity or wrinkles containing lactic acid bacteria fermented rice germ extract as an active ingredient.
A cosmetic composition for improving skin moisturization containing lactic acid bacteria fermented rice germ extract as an active ingredient.